Development and validation of RP-HPLC method for the simultaneous estimation of sitagliptin phosphate and dapagliflozin in bulk and marketed formulations with forced degradation studies
Keywords:
Sitagliptin Phosphate, Dapagliflozin, RP-HPLC, Method Validation, Forced Degradation, Linearity, Stability StudiesAbstract
Introduction: The present study focuses on the development and validation of novel RP-HPLC method for the simultaneous estimation of Sitagliptin phosphate and Dapagliflozin in bulk drugs and marketed formulations. Method validation was carried out in accordance with International Council for Harmonisation (ICH) guidelines, assessing critical parameters such as accuracy, linearity, precision, specificity, limit of detection (LOD), and limit of quantification (LOQ)[1,2]. Additionally, forced degradation studies were performed under acidic, alkaline, oxidative, photolytic, and thermal conditions to evaluate the stability-indicating capability of the developed methods. These validated analytical tools are expected to offer reliable and reproducible results for routine quality assurance, regulatory compliance, and stability testing of combination antidiabetic formulations.RP-HPLC methods for their estimation, along with assessing their stability through forced degradation studies.
Materials and Methods: The RP-HPLC method utilized a reversed-phase C18 column with a mobile phase composition optimized to achieve sharp, well-resolved peaks. Validation of method was performed according to ICH Q2(R1) guidelines, assessing parameters such as linearity, precision, accuracy, limit of detection (LOD), limit of quantitation (LOQ), ruggedness, and robustness. Forced degradation studies were conducted under various stress conditions including acidic, alkaline, oxidative, photolytic, and thermal environments.
Results: The RP-HPLC method showed distinct retention times of 5.473 minutes for Sitagliptin and 8.611 minutes for Dapagliflozin. Both methods demonstrated high accuracy and precision with acceptable %RSD values (<2%). LOD and LOQ values confirmed the sensitivity of the methods. Among all stress conditions, the maximum degradation was observed under alkaline conditions, indicating that both drugs are less stable in basic environments.
Conclusion: The developed RP-HPLC method is simple, sensitive, and reliable for the simultaneous estimation of Sitagliptin Phosphate and Dapagliflozin in bulk and marketed formulations. Their successful validation and application to forced degradation studies underscore their suitability for routine quality control, formulation analysis, and stability testing in pharmaceutical industries
Downloads
References
International Conference on Harmonisation (ICH). ICH Q2(R1) Validation of Analytical Procedures: Text and Methodology. Geneva: ICH; 2005.
Chatwal GR, Anand KS. Instrumental Methods of Chemical Analysis. 5th ed. Mumbai: Himalaya Publishing House; 2014.
International Conference on Harmonisation (ICH). Stability Testing of New Drug Substances and Products Q1A(R2). Geneva: ICH; 2003.
Pawar AR, Patil SR. RP-HPLC method development and validation for simultaneous estimation of Sitagliptin and Dapagliflozin in tablet dosage form. Asian J Pharm Clin Res. 2018;11(3):208-11.
World Health Organization. International Pharmacopoeia. General Methods and Stability Studies. Geneva: WHO; 2022. Available from: https://www.who.int/publications
Singh S, Bakshi M. Guidance on conduct of stress tests to determine inherent stability of drugs. Pharm Technol Online. 2000;24(2):1-14.
Skoog DA, Holler FJ, Crouch SR. Principles of Instrumental Analysis. 7th ed. Boston: Cengage Learning; 2017.
Meyer VR. Practical High-Performance Liquid Chromatography. 5th ed. Chichester: Wiley; 2010.
Tiwari G, Tiwari R, Rai AK. Review on analytical method development and validation. Int J Pharm Life Sci. 2010;1(5):310–7.
Sahu PK, Ramisetti NR, Cecchi T, Swain S, Patro CS, Panda J. An overview of experimental designs in HPLC method development and validation. J Pharm Biomed Anal. 2018;147:590–611.
International Conference on Harmonisation (ICH). Q1B: Photostability Testing of New Drug Substances and Products. Geneva: ICH; 1996.
Naik AA, Pawar D. UV and HPLC method development and validation for simultaneous estimation of Sitagliptin phosphate and Dapagliflozin. Int J Pharm ChemBiol Sci. 2017;7(2):70–5.
Rane VP, Patil KR, Sangshetti JN, Yeole RD, Shinde DB. Stability-indicating LC method for the determination of Sitagliptin in bulk drug and pharmaceutical dosage form. Sci Pharm. 2010;78(3):477–92. doi:10.3797/scipharm.0910-15
Dhalwal K, Shinde VM, Namdeo AG. Stability testing of phytopharmaceuticals: A review. Pharmacogn Rev. 2008;2(4):215–8.
Vankalapati KR, Alegete P, Boodida S. Stability-indicating HPLC method development and validation for simultaneous estimation of metformin, dapagliflozin, and saxagliptin in bulk drug and pharmaceutical dosage form. Biomed Chromatogr. 2022;36(7):e5384. https://doi.org/10.1002/bmc.5384
Gurrala S, Shivaraj C, Anumolu PD, Saraf G. Multivariate optimization of liquid chromatographic conditions for simultaneous analysis of dapagliflozin and saxagliptin in bulk and pharmaceutical dosage form. Future J Pharm Sci. 2021;7(1):85. https://doi.org/10.1186/s43094-021-00229-z
Acharya H, Kotadiya R. Stability-indicating HPLC for remogliflozin, vildagliptin, and metformin: Method development, validation, and greenness. Ann Pharm Fr. 2024;82(6):1071–81. https://doi.org/10.1016/j.pharma.2024.06.006
Vankalapati KR, Alegete P, Boodida S. Stability-indicating ultra performance liquid chromatography method development and validation for simultaneous estimation of metformin, linagliptin, and empagliflozin in bulk and pharmaceutical dosage form. Biomed Chromatogr. 2021;35(4):e5019. https://doi.org/10.1002/bmc.5019
Sanagapati M, Dhanalakshmi K, Nagarjuna R. Novel univariate spectrophotometric determination of the recently approved anti-diabetic agent dapagliflozin in bulk and pharmaceutical formulations. SpectrochimActa A MolBiomolSpectrosc. 2019;213:86–90. https://doi.org/10.1016/j.saa.2019.01.056
Vankalapati KR, Alegete P, Boodida S. Development and validation of simplified RP-HPLC method for determination of darunavir in bulk and pharmaceutical dosage form. Mater Today Proc. 2021;47:394–9. https://doi.org/10.1016/j.matpr.2021.04.097
Bansal G, Singh M, Jindal K. Identification, isolation and characterization of potential process impurities and degradation products of glimepiride: Development of validated stability-indicating method. J Pharm Biomed Anal. 2015;115:312–22. https://doi.org/10.1016/j.jpba.2015.07.015
Kumar V, Chawla G. Simple and rapid LC-MS/MS method for determination of sitagliptin in human plasma and its application to a pharmacokinetic study. J Pharm Biomed Anal. 2020;189:113426. https://doi.org/10.1016/j.jpba.2020.113426
El-Bagary RI, Elkady EF, Ayoub BM. Second-derivative synchronous fluorimetry and time-programmed flow injection technique for simultaneous determination of vildagliptin and saxagliptin. J Pharm Biomed Anal. 2015;111:156–63. https://doi.org/10.1016/j.jpba.2015.03.011
Bansal G, Singh M, Jindal K. RP-HPLC method for simultaneous estimation of bisoprolol fumarate and hydrochlorothiazide in tablet formulation. J Pharm Biomed Anal. 2009;50(5):887–90. https://doi.org/10.1016/j.jpba.2009.06.019
Gurrala S, Shivaraj C, Anumolu PD. Multivariate optimization of liquid chromatographic conditions for simultaneous quantification of dapagliflozin and saxagliptin: Application to in vitro dissolution and stability studies. Future J Pharm Sci. 2021;7(1):85. https://doi.org/10.1186/s43094-021-00229-z
Sanagapati M, Dhanalakshmi K, Reddy NV. Novel univariate spectrophotometric determination of the recently approved antidiabetic agent dapagliflozin in bulk and pharmaceutical formulations. SpectrochimActa A MolBiomolSpectrosc. 2019;223:117353. https://doi.org/10.1016/j.saa.2019.117353
Abdelrahman AE, Maher HM, Alzoman NZ. HPTLC method for the determination of metformin hydrochloride, saxagliptin hydrochloride, and dapagliflozin in pharmaceuticals. Curr Anal Chem. 2020;16(5):609–19. https://doi.org/10.2174/1573411015666190131123029
Shakoor A, Ahmed M, Ikram R, Hussain S, Tahir A, Jan BM, et al. Stability-indicating RP-HPLC method for simultaneous determination of metformin hydrochloride and vildagliptin in tablet and biological samples. ActaChromatogr. 2020;32(1):39–43. https://doi.org/10.1556/1326.2019.00677
Downloads
Published
How to Cite
Issue
Section
License
This work is licensed under a Creative Commons Attribution 4.0 International License.
You are free to:
- Share — copy and redistribute the material in any medium or format
- Adapt — remix, transform, and build upon the material for any purpose, even commercially.
Terms:
- Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
- No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.
